MCL1激活MAPK-MEK信号通路促进食管癌细胞EC9706增殖  

MCL1 Promotes Cell Proliferation of Esophageal Cancer Cell EC9706 Through Activating MAPK-MEK Signaling Pathway

作　　者：崔艳丽[1] 何利珍[1] CUI Yanli;HE Lizhen(Laboratory of Second People’s Hospital of Jiaozuo City,He’nan Province 454000)

机构地区：[1]河南省焦作市第二人民医院检验科,454000

出　　处：《医学理论与实践》2021年第10期1625-1629,共5页The Journal of Medical Theory and Practice

摘　　要：目的:探讨食管癌中MCL1表达变化及其调控食管癌细胞EC9706增殖的机制。方法:对2018年期间收集的12份食管癌组织及其癌周组织进行相对定量检测MCL1基因表达情况。构建表达MCL1的真核表达载体pCDNA3.1 H-MCL1-myc-His;将EC9706细胞设置转染空载体pCDNA3.1-myc-His的对照组、转染pCDNA3.1-MCL1-myc-His的实验组,细胞计数检测MCL1对细胞数量的影响,流式细胞仪检测细胞周期中各时期细胞比例变化,划痕实验检测对细胞迁移能力的影响。并以Western blot检测MCL1对MEK1磷酸化的影响,qRT-PCR检测MCL1对MAPK-MEK下游基因C-MYC、C-FOS、CCND1激活情况。结果:成功构建了表达MCL1的真核表达载体。和对照组相比,超表达MCL1后能够调控细胞周期改变,显著促进EC9706细胞数量增加,尤其在超表达MCL148h后,差异显著(P≤0.05);流式细胞仪检测细胞周期变化时也发现超表达MCL1后能够增加S+G2期的细胞比例;细胞划痕实验显示MCL1能够提高细胞迁移能力。检测到MCL1能够提高p-MEK1含量,并激活MAPK-MEK下游的靶基因C-MYC、C-FOS、CCND1的转录,差异显著(P≤0.05)。结论:MCL1在食管癌中普遍高表达,并且能够通过激活MAPK-MEK信号通路促进食管癌细胞EC9706增殖,或许可以作为一个鉴别食管癌的候选基因。Objective:To investigate the expression changes of MCL1 in esophageal cancer tissues and the mechanisms for regulating EC9706 cell proliferation.Methods:A total of 12 cases of esophageal cancer tissue specimens(cancer tissue)collected of 2018,and qRT-PCR technique was used to detect the expression of MCL1 in cancer tissue and in cancer surrounding tissues.The eukaryotic expression vector pCDNA3.1-MCL1-myc-His were constructed.The pCDNA3.1-myc-His vector as control,and transfected vectors into EC9706 cells,defect the reproductive ability of each group by counting cells and flow cytometry.The migration of the MCL1 over expressed cells was examined by cell scratch test.Western blot and qRT-PCR test to the activation of MAPK-MEK signal transduction pathway by MCL1.Results:The pCDNA3.1-MCL1-myc-His plasmid was successfully constructed.Compared with control group,overexpressed MCL1 increase the cell amount significantly(P≤0.05),and increases the S+G2 phase ratio.Cell scratch test showed MCL1 promotes migration.Western blot results shows MCL1 increased the p-MEK1 protein,and significantly increased the downstream gene(C-MYC,C-FOS,CCND1)expression of MAPK-MEK signal transduction pathway.Conclusion:MCL1 protein is over expressed in esophageal carcinoma,and over expression MCL1 can mediates activation of the MAPK-MEK transcription pathway then benefit to EC9706 cell proliferation,which may suggest that high expression of MCL1 gene could be one of the causes in the carcinogenesis of human esophageal carcinoma.

关 键 词：食管癌 MAPK-MEK信号通路 MCL1 细胞增殖 

分 类 号：R735.1[医药卫生—肿瘤]